Crystalline forms of a dipeptidyl peptidase-IV inhibitors

ABSTRACT

Novel crystalline forms of (2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine are potent inhibitors of dipeptidyl peptidase-IV and are useful for the treatment of non-insulin dependent (Type 2) diabetes mellitus. The invention also relates to pharmaceutical compositions containing these novel forms, processes to prepare these forms and their pharmaceutical compositions as well as uses thereof for the treatment of Type 2 diabetes.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is continuation of U.S. patent application Ser. No.14/118, 998, filed 20 Nov. 2013, which is the National Stage ofInternational Application No. PCT/US2012/043922, filed 25 Jun. 2012,which claims the benefit under 35 U.S.C. 119(e) of U.S. ProvisionalApplication No. 61/502,497, filed 29 Jun. 2011.

FIELD OF THE INVENTION

The present invention relates to novel crystalline forms of a dipeptidylpeptidase-IV inhibitor. More particularly, the invention relates tonovel crystalline forms of(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine,which is a potent, long acting inhibitor of dipeptidyl peptidase-IV.These novel crystalline forms, are useful for the treatment andprevention of diseases and conditions for which an inhibitor ofdipeptidyl peptidase-IV is indicated, in particular Type 2 diabetes,obesity, and high blood pressure. The invention further concernspharmaceutical compositions comprising the novel crystalline forms ofthe present invention useful to treat Type 2 diabetes, obesity, and highblood pressure as well as processes for the preparation of such formsand their pharmaceutical compositions.

BACKGROUND OF THE INVENTION

Inhibition of dipeptidyl peptidase-IV (DP-IV), an enzyme thatinactivates both glucose-dependent insulinotropic peptide (GIP) andglucagon-like peptide 1 (GLP-1), represents a novel approach to thetreatment and prevention of Type 2 diabetes, also known as non-insulindependent diabetes mellitus (NIDDM). The therapeutic potential of DP-IVinhibitors for the treatment of Type 2 diabetes has been reviewed: C. F.Deacon and J. J. Holst, “Dipeptidyl peptidase IV inhibition as anapproach to the treatment and prevention of Type 2 diabetes: ahistorical perspective,” Biochem. Biophys. Res. Commun., 294: 1-4(2000); K. Augustyns, et al., “Dipeptidyl peptidase IV inhibitors as newtherapeutic agents for the treatment of Type 2 diabetes,” Expert. Opin.Ther. Patents, 13: 499-510 (2003); D. J. Drucker, “Therapeutic potentialof dipeptidyl peptidase IV inhibitors for the treatment of Type 2diabetes,” Expert Opin. Investig. Drugs, 12: 87-100 (2003); and M. A.Nauck et al., “Incretins and Their Analogues as New Antidiabetic Drugs,”Drug News Perspect., 16: 413-422 (2003).

WO 2010/056708 (published 20 May 2010), assigned to Merck & Co.,describes a class of aminotetrahydropyrans, which are potent inhibitorsof DP-IV and therefore useful for the treatment of Type 2 diabetes.Specifically disclosed in WO 2010/056708 is(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.

However, the applicants have now discovered novel crystalline forms of(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Compound I).

SUMMARY OF THE INVENTION

The present invention is concerned with novel crystalline forms of thedipeptidyl peptidase-IV (DP-IV) inhibitor(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Compound I). Certain crystalline forms, have advantages in thepreparation of pharmaceutical compositions of(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine,such as ease of processing and crystallisation, handling, stability tostress and dosing. In particular, they exhibit improved physicochemicalproperties, such as stability to stress, rendering them particularlysuitable for the manufacture of various pharmaceutical dosage forms. Theinvention also concerns pharmaceutical compositions containing the novelforms thereof, as well as methods for using them as DP-IV inhibitors, inparticular for the prevention or treatment of Type 2 diabetes, obesity,and high blood pressure. In certain embodiments, described herein arepharmaceutical compositions comprising crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amineand a pharmaceutically acceptable carrier.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a X-ray diffraction pattern of crystalline Form I of CompoundI.

FIG. 2 is a thermogravimetric analysis (TGA) curve of crystalline Form Iof Compound I.

FIG. 3 is a differential scanning calorimetry (DSC) curve of crystallineForm I of Compound I.

FIG. 4 is a solid state NMR spectra of crystalline Form I of Compound I.

FIG. 5 is an IR spectra of crystalline Form II of Compound I.

FIG. 6 is a X-ray diffraction pattern of crystalline Form II of CompoundI.

FIG. 7 is a thermogravimetric analysis (TGA) curve of crystalline FormII of Compound I.

FIG. 8 is a differential scanning calorimetry (DSC) curve of crystallineForm II of Compound I.

FIG. 9 is a solid state NMR spectra of crystalline Form II of CompoundI.

FIG. 10 is an IR spectra of crystalline Form II of Compound I.

FIG. 11 is a X-ray diffraction pattern of crystalline Form III ofCompound I.

FIG. 12 is a thermogravimetric analysis (TGA) curve of crystalline FormIII of Compound I.

FIG. 13 is a differential scanning calorimetry (DSC) curve ofcrystalline Form III of Compound I.

FIG. 14 is a X-ray diffraction pattern of crystalline Form IV ofCompound I.

FIG. 15 is a thermogravimetric analysis (TGA) curve of crystalline FormIV of Compound I.

FIG. 16 is a differential scanning calorimetry (DSC) curve ofcrystalline Form IV of Compound I.

DETAILED DESCRIPTION OF THE INVENTION

This invention relates to crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amineof Compound I:

Unless a specific form designation is given, the term “crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-aminerefers to all crystalline forms of(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-aminedescribed herein. The crystalline forms described herein exist as theanhydrous free base of(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.

One embodiment of the crystalline forms described herein is(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Form I). Form I is further described below.

Another embodiment of the crystalline forms described herein is(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Form II). Form II is further described below.

Still another embodiment of the crystalline forms described herein is(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Form III). Form III is further described below.

Yet another embodiment of the crystalline forms described herein is(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Form IV). Form IV is further described below.

A further embodiment of the present invention provides a particular drugsubstance that comprises at least one of the crystalline forms describedherein. By “drug substance” is meant the active pharmaceuticalingredient. The amount of crystalline form in the drug substance can bequantified by the use of physical methods such as X-ray powderdiffraction, solid-state fluorine-19 magic-angle spinning (MAS) nuclearmagnetic resonance spectroscopy, solid-state carbon-13cross-polarization magic-angle spinning (CPMAS) nuclear magneticresonance spectroscopy, solid state Fourier-transform infraredspectroscopy, and Raman spectroscopy.

In a class of this embodiment, the crystalline form of the presentinvention is present in about 5% to about 100% by weight of the drugsubstance. In a second class of this embodiment, the crystalline form ofthe present invention is present in about 10% to about 100% by weight ofthe drug substance. In a third class of this embodiment, the crystallineform of the present invention is present in about 25% to about 100% byweight of the drug substance. In a fourth class of this embodiment, thecrystalline form of the present invention is present in about 50% toabout 100% by weight of the drug substance. In a fifth class of thisembodiment, the crystalline form of the present invention is present inabout 75% to about 100% by weight of the drug substance. In a sixthclass of this embodiment, substantially all of the drug substance is thecrystalline form of the present invention, i.e., the drug substance issubstantially phase pure crystalline.

In another class of this embodiment, at least 5% by weight of the drugsubstance is the crystalline form of the present invention. In a yetanother class of this embodiment, at least 10% by weight of the drugsubstance is the crystalline form of the present invention. In a stillanother class of this embodiment, at least 15% by weight of the drugsubstance is the crystalline form of the present invention. In anotherclass of this embodiment, at least 20% by weight of the drug substanceis the crystalline form of the present invention. In yet another classof this embodiment, at least 25% by weight of the drug substance is thecrystalline form of the present invention. In still another class ofthis embodiment, at least 30% by weight of the drug substance is thecrystalline form of the present invention. In another class of thisembodiment, at least 35% by weight of the drug substance is thecrystalline form of the present invention. In a yet another class ofthis embodiment, at least 40% by weight of the drug substance is thecrystalline form of the present invention. In a still another class ofthis embodiment, at least 45% by weight of the drug substance is thecrystalline form of the present invention. In another class of thisembodiment, at least 50% by weight of the drug substance is thecrystalline form of the present invention. In yet another class of thisembodiment, at least 55% by weight of the drug substance is thecrystalline form of the present invention. In still another class ofthis embodiment, at least 60% by weight of the drug substance is thecrystalline form of the present invention. In another class of thisembodiment, at least 65% by weight of the drug substance is thecrystalline form of the present invention. In a yet another class ofthis embodiment, at least 70% by weight of the drug substance is thecrystalline form of the present invention. In a still another class ofthis embodiment, at least 75% by weight of the drug substance is thecrystalline form of the present invention. In another class of thisembodiment, at least 80% by weight of the drug substance is thecrystalline form of the present invention. In yet another class of thisembodiment, at least 85% by weight of the drug substance is thecrystalline form of the present invention. In still another class ofthis embodiment, at least 90% by weight of the drug substance is thecrystalline form of the present invention. In another class of thisembodiment, at least 95% by weight of the drug substance is thecrystalline form of the present invention. In a yet another class ofthis embodiment, at least 100% by weight of the drug substance is thecrystalline form of the present invention.

The crystalline forms of the present invention exhibit pharmaceuticaladvantages over the amorphous free base of Compound I as described in WO2010/056708 in the preparation of a pharmaceutical drug productcontaining the pharmacologically active ingredient. In particular, theenhanced chemical and physical stability of the crystalline formsconstitute advantageous properties in the preparation of solidpharmaceutical dosage forms containing the pharmacologically activeingredient.

The crystalline forms of the present invention, which exhibit longacting, potent DP-IV inhibitory properties, are particularly useful forthe prevention or treatment of Type 2 diabetes, obesity, and high bloodpressure.

Another aspect of the present invention provides a method for theprevention or treatment of clinical conditions for which an inhibitor ofDP-IV is indicated, which method comprises administering to a patient inneed of such prevention or treatment a prophylactically ortherapeutically effective amount of a crystalline form of the presentinvention, or a hydrate thereof. Such clinical conditions includediabetes, in particular Type 2 diabetes, hyperglycemia, insulinresistance, and obesity.

The present invention also provides for the use of a crystalline form ofCompound I of the present invention for the prevention or treatment in amammal of clinical conditions for which an inhibitor of DP-IV isindicated, in particular Type 2 diabetes, hyperglycemia, insulinresistance, and obesity.

The present invention also provides for the use of a crystalline form ofCompound I of the present invention for the manufacture of a medicamentfor the prevention or treatment in a mammal of clinical conditions forwhich an inhibitor of DP-IV is indicated, in particular Type 2 diabetes,hyperglycemia, insulin resistance, and obesity.

The present invention also provides pharmaceutical compositionscomprising a crystalline form described herein, in association with oneor more pharmaceutically acceptable carriers or excipients. In oneembodiment the pharmaceutical composition comprises a therapeuticallyeffective amount of the active pharmaceutical ingredient in admixturewith pharmaceutically acceptable excipients wherein the activepharmaceutical ingredient comprises a detectable amount of a crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.

In a second embodiment the pharmaceutical composition comprises atherapeutically effective amount of the active pharmaceutical ingredientin an admixture with pharmaceutically acceptable excipients wherein theactive pharmaceutical ingredient comprises about 1% to about 100% byweight of crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.In a class of this second embodiment, the active pharmaceuticalingredient in such compositions comprises about 5% to about 100% byweight of crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.In a second class of this embodiment, the active pharmaceuticalingredient in such compositions comprises about 10% to about 100% byweight of crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.In a third class of this embodiment, the active pharmaceuticalingredient in such compositions comprises about 25% to about 100% byweight of crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.In a fourth class of this embodiment, the active pharmaceuticalingredient in such compositions comprises about 50% to about 100% byweight of crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.

In a third embodiment the pharmaceutical composition comprises atherapeutically effective amount of the active pharmaceutical ingredientin an admixture with pharmaceutically acceptable excipients wherein theactive pharmaceutical ingredient comprises at least 1% by weight ofcrystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.In a class of this second embodiment, the active pharmaceuticalingredient in such compositions comprises about 5% by weight ofcrystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.In a second class of this embodiment, the active pharmaceuticalingredient in such compositions comprises at least 10% by weight ofcrystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.In a third class of this embodiment, the active pharmaceuticalingredient in such compositions comprises at least 25% by weight ofcrystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.In a fourth class of this embodiment, the active pharmaceuticalingredient in such compositions comprises at least 50% by weight ofcrystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine.

The compositions in accordance with the invention are suitably in unitdosage forms such as tablets, pills, capsules, powders, granules,sterile solutions or suspensions, metered aerosol or liquid sprays,drops, ampoules, auto-injector devices or suppositories. Thecompositions are intended for oral, parenteral, intranasal, sublingual,or rectal administration, or for administration by inhalation orinsufflation. Formulation of the compositions according to the inventioncan conveniently be effected by methods known from the art, for example,as described in Remington's Pharmaceutical Sciences, 17^(th) ed., 1995.

The dosage regimen is selected in accordance with a variety of factorsincluding type, species, age, weight, sex and medical condition of thepatient; the severity of the condition to be treated; the route ofadministration; and the renal and hepatic function of the patient. Anordinarily skilled physician, veterinarian, or clinician can readilydetermine and prescribe the effective amount of the drug required toprevent, counter or arrest the progress of the condition.

Oral dosages of the present invention, when used for the indicatedeffects, will range between about 0.01 mg per kg of body weight per day(mg/kg/day) to about 100 mg/kg/day, preferably 0.01 to 10 mg/kg/day, andmost preferably 0.1 to 5.0 mg/kg/day. For oral administration, thecompositions are preferably provided in the form of tablets containing0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100 and 500milligrams of the active ingredient for the symptomatic adjustment ofthe dosage to the patient to be treated. A medicament typically containsfrom about 0.01 mg to about 500 mg of the active ingredient, preferably,from about 1 mg to about 200 mg of active ingredient. Intravenously, themost preferred doses will range from about 0.1 to about 10 mg/kg/minuteduring a constant rate infusion. The crystalline forms of the presentinvention may be administered in a single daily dose, or the total dailydosage may be administered in divided doses of two, three or four timesdaily. However,(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amineis a long acting DPP-IV inhibitor. Advantageously, the crystalline formsof the present invention may be administered in a single weekly dose.

Furthermore, the crystalline forms of the present invention can beadministered in intranasal form via topical use of suitable intranasalvehicles, or via transdermal routes, using those forms of transdermalskin patches well known to those of ordinary skill in the art. To beadministered in the form of a transdermal delivery system, the dosageadministration will, of course, be continuous rather than intermittentthroughout the dosage regimen.

In the methods of the present invention, the crystalline forms describedherein can form the active pharmaceutical ingredient, and are typicallyadministered in admixture with suitable pharmaceutical diluents,excipients or carriers (collectively referred to herein as ‘carrier’materials) suitably selected with respect to the intended form ofadministration, that is, oral tablets, capsules, elixirs, syrups and thelike, and consistent with conventional pharmaceutical practices.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic, pharmaceutically acceptable, inert carrier such as lactose,starch, sucrose, glucose, methyl cellulose, magnesium stearate,dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like;for oral administration in liquid form, the oral drug component can becombined with any oral, non-toxic, pharmaceutically acceptable inertcarrier such as ethanol, glycerol, water and the like. Moreover, whendesired or necessary, suitable binders, lubricants, disintegratingagents and coloring agents can also be incorporated into the mixture.Suitable binders include starch, gelatin, natural sugars such as glucoseor beta-lactose, corn sweeteners, natural and synthetic gums such asacacia, tragacanth or sodium alginate, carboxymethylcellulose,polyethylene glycol, waxes and the like. Lubricants used in these dosageforms include sodium oleate, sodium stearate, magnesium stearate, sodiumbenzoate, sodium acetate, sodium chloride and the like. Disintegratorsinclude, without limitation, starch, methyl cellulose, agar, bentonite,xanthan gum and the like.

The crystalline forms of Compound I of the present invention have beenfound to possess a relatively high solubility in water (about 2 mg/ml),rendering them especially amenable to the preparation of formulations,in particular intranasal and intravenous formulations, which requirerelatively concentrated aqueous solutions of active pharmaceuticalingredient.

In a still further aspect, the present invention provides a method forthe treatment and/or prevention of clinical conditions for which a DP-IVinhibitor is indicated, which method comprises administering to apatient in need of such prevention or treatment a prophylactically ortherapeutically effective amount of a crystalline form of Compound I asdefined above in combination with another agent useful for the treatmentof Type 2 diabetes, obesity, and high blood pressure.

Compounds described herein may exist as tautomers such as keto-enoltautomers. The individual tautomers as well as mixtures thereof areencompassed with compounds of structural formula I.

The term “% enantiomeric excess” (abbreviated “ee”) shall mean the %major enantiomer less the % minor enantiomer. Thus, a 70% enantiomericexcess corresponds to formation of 85% of one enantiomer and 15% of theother. The term “enantiomeric excess” is synonymous with the term“optical purity.”

Compound I can be made by the following methods:

INTERMEDIATE 1

tert-Butyl[(2R,3S)-5-oxo-2-(2,5-difluorophenyl)tetrahydro-2H-pyran-3-yl]carbamateStep A: tert-Butyl(1-[methoxy(methyl)amino]-1-oxopent-4-yn-2-yl)carbamate

To an inerted vessel was charged N,N-diphenyl glycine ethyl ester(105.45 kg, 394.5 mol), tetrabutyl ammonium bromide (14 kg, 43.4 mol),and propargyl benzenesulfonate (94.45 kg, 481 mol) followed by MTBE (750kg). Then cesium carbonate (fine mesh grade, 390 kg, 1197 mol) was addedand the reaction stirred at 50-60° C. for 1 day. The batch was thencooled to 0-5° C. and water (422 kg) was slowly added. Next, tert-butylmethyl ether (170 kg) was added and the batch concentrated to 473-578 L.Then, 462 kg HCl solution (43 kg conc. HCl in 420 kg water) was added toreach a pH=1-2 below room temperature. After 7 h of stirring, the pH was1.5 and the organic layer was separated and discarded.

The aqueous layer was then cooled to 5-10° C. and 28% aqueous NaOH (151kg) was added slowly until the pH was 13. Then, a solution of Boc₂O (136kg, 624 mol in 243 kg of tert-butyl methyl ether) was added at 5-10° C.The solution was then stirred at room temperature for 4 h (pH=8) and 17%aqueous NaOH (126 kg) was slowly added followed by more Boc₂O solution(30.7 kg, 141 mol in 60 kg tert-butyl methyl ether). The solution wasthen stirred at room temperature for 4 h (pH=9) and 17% aqueous NaOH (98kg) was slowly added (pH=13) and stirred an additional 12 h (pH-10)followed by more Boc₂O (11 kg, 50 mol). After 4 h of stirring at roomtemperature, the layers were separated (retained aqueous) and theorganics extracted with 3% aqueous NaOH (136 kg). The aqueous layerswere combined and added to tert-butyl methyl ether (338 kg). Then,aqueous 17% HCl (362 kg) was added until pH=2. The layers were separatedand the aqueous extracted with tert-butyl methyl ether (420 kg). Thecombined organics were washed with 10% brine (139 kg), dried withNa₂SO₄, filtered, and concentrated to 105-158 L. Constant volumedistillation with tert-butyl methyl ether continued until KF=0.4%.

Carbonyldiimidazole (90 kg, 548 mol) was added to this solution andstirred for 2 h at room temperature. Then (MeO)MeNH₂Cl (48 kg, 492 mol)was added and the reaction stirred for 6 h. The batch was then cooled to0-5° C. and water (80 kg) was added. The batch was then seeded with 100g seed and water (450 kg) was added. The slurry was stirred at 0-5° C.for 3 h and then filtered. The cake was dried under vacuum at 45-60° C.for 2 days to give tert-butyl(1-[methoxy(methyl)amino]-1-oxopent-4-yn-2-yl)carbamate.

Step B: tert-Butyl [1-(2,5-difluorophenyl)-1-oxopent-4-yn-2-yl]carbamate

An inerted vessel was charged dichloromethane (866 kg) and cooled to −20to −10° C. Then iso-propylmagnesium chloride solution in THF (2M, 326.1kg, 669 mol) was slowly added followed by 1-bromo-2,5-difluorobenzene(120.1 kg, 622 mol). After 2 h at this temperature, an additional chargeof iso-propylmagnesium chloride in THF solution was slowly added (2M,58.65 kg, 121 mol) and the reaction aged 1 h. Then, a drop-wise additionof a dichloromethane solution of tert-butyl(1-[methoxy(methyl)amino]-1-oxopent-4-yn-2-yl)carbamate (70.8 kg, 276mol in 292 kg dichloromethane) was conducted over 2 h at −20 to −20° C.The mixture was then warmed to room temperature and stirred for 10 h.The reaction was then slowly reverse quenched into aqueous ammoniumchloride (175.6 kg in 1550 kg of water) at 5-10° C. The solution pH wasthen adjusted to ˜7 by adding 68 kg of con. HCl. The layers were thenseparated and the aqueous extracted with dichloromethane (414 kg). Thecombined organics were then dried with Na₂SO₄, filtered, treated withactivated carbon (10 kg), filtered, and concentrated to 71-141 L. Aconstant volume (71-141 L) vacuum distillation solvent switch ton-heptane was then performed to crystallize the product. The slurry wasthen cooled to 0° C. and stirred 2 h. The slurry was filtered and thecake washed with n-heptane, 2-propanol, and then water. The solids weredried under vacuum at 40-50° C. overnight to give tert-butyl[1-(2,5-difluorophenyl)-1-oxopent-4-yn-2-yl]carbamate.

Step C: tert-Butyl[(1S,2S)-1-(2,5-difluorophenyl)-1-hydroxypent-4-yn-2-yl]carbamate

To a stirred vessel under nitrogen sweep was charged tert-butyl[1-(2,5-difluorophenyl)-1-oxopent-4-yn-2-yl]carbamate (35.0 kg, 113mol), 1,4-diazabicyclo[2.2.2]octane (38.0 kg, 339 mol), and THF (465kg). After dissolution,chloro{[(1R,2R)-(+2-amino-1,2-diphenylethyl](pentafluorophenylsulfonyl)amido}-(p-cymene)ruthenium (II) (410 g, 576 mmol) was added. The vessel was vacuumsparged and back-filled with nitrogen three times. Then, formic acid(26.7 kg, 580 mol) was added and the reaction heated to 45° C.overnight.

The mixture was then concentrated under vacuum to 210-280 L andtert-butyl methyl ether was then added (210 kg). After cooling to 0-10°C., 0.4% aqueous HCl was added (52 kg) until pH=4-6. After agitation andseparation of the layers, the aqueous was extracted again withtert-butyl methyl ether (87 kg). The combined organics were then washedwith 4% aq. NaHCO₃ (291 kg), and then brine (216 kg). The resultingorganics were dried over Na₂SO₄, filtered through a plug of silica, andconcentrated to 70-105 L. Then, tert-butyl methyl ether (132 kg) wasadded, followed by further batch concentration until KF=0.1%. Next, DMF(133 kg) was added and the batch was further concentrated to 70-105 L.The resulting DMF solution was 165.6 kg containing 19.4% tert-butyl[(1S,2S)-1-(2,5-difluorophenyl)-1-hydroxypent-4-yn-2-yl]carbamate (8.1/1diastereomeric ratio and 97.9% ee).

Step D: tert-Butyl[(1S,2R)-1-(2,5-difluorophenyl)-1-hydroxypent-4-yn-2-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step C.

Step E: tert-Butyl[(1R,2R)-1-(2,5-difluorophenyl)-1-hydroxypent-4-yn-2-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step D.

Step F: tert-Butyl[(1R,2S)-1-(2,5-difluorophenyl)-1-hydroxypent-4-yn-2-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step E.

Step G: tert-Butyl[(2R,3S)-2-(2,5-difluorophenyl)-3,4-dihydro-2H-pyran-3-yl]carbamate

To a 165.6 kg solution of tert-butyl[(1S,2S)-1-(2,5-difluorophenyl)-1-hydroxypent-4-yn-2-yl]carbamate (19.4w/w % in DMF, 103 mol) was added DMF (70 kg),1-hydroxypyrrolidine-2,5-dione (5.95 kg, 51 mol), tetrabutylammoniumhexafluorophosphate (5.20 kg, 13 mol), and NaHCO₃ (4.50 kg, 54 mol). Theresulting reaction mixture was vacuum sparged with a nitrogen back-fillthree times and then stirred for 30-40 min. Then,chloro(cyclopentadienyl)bis(triphenylphosphine) ruthenium (II) (823 g,1.13 mol) and triphenylphosphine (892 g, 3.40 mol) was added and thereaction was vacuum purged with nitrogen back-filling three times. Thereaction was then heated to 75-85° C. overnight. To complete thereaction, additional chloro(cyclopentadienyl)bis(triphenylphosphine)ruthenium (II) (826 g, 1.14 mol) and triphenylphosphine (892 g, 3.40mol) was added and the reaction heated at 75-85° C. an additional 12-16h.

After cooling to room temperature, water (250 kg) and tert-butyl methylether (210 kg) was added. After agitation, the layers were separated andthe resulting aqueous layer was extracted with tert-butyl methyl ether(2×150 kg). The combined organics were washed with brine (4×220 kg). Theorganics were then dried with Na₂SO₄, filtered, and concentrated. Thecrude was passed through a plug of silica with tert-butyl methyl etherand n-heptane. The resulting solution was then solvent switched byvacuum distillation and feeding n-heptane to a slurry of 64-128 L inn-heptane. This slurry was heated to dissolve at 90-110° C. This wasthen cooled over 2-3 h to 0-10° C. The slurry was then filtered and theresulting wet cake dried at 40-50° C. and vacuum to give tert-butyl[(2R,3S)-2-(2,5-difluorophenyl)-3,4-dihydro-2H-pyran-3-yl]carbamate.

Step H: tert-Butyl[(2R,3R)-2-(2,5-difluorophenyl)-3,4-dihydro-2H-pyran-3-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step G.

Step I: tert-Butyl[(2S,3S)-2-(2,5-difluorophenyl)-3,4-dihydro-2H-pyran-3-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step H.

Step J: tert-Butyl[(2S,3R)-2-(2,5-difluorophenyl)-3,4-dihydro-2H-pyran-3-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step I.

Step K: tert-Butyl[(2R,3S)-2-(2,5-difluorophenyl)-5-hydroxytetrahydro-2H-pyran-3-yl]carbamate

To 64.0 kg (206 mol) of tert-butyl[(2R,3S)-2-(2,5-difluorophenyl)-3,4-dihydro-2H-pyran-3-yl]carbamate in astirred vessel was added tert-butyl methyl ether (500 kg). Afterdissolving, the solution was cooled to 0-5° C. and 10M borane-dimethylsulfide complex solution was added (39 kg, 515 mol). After 1-3 h ofstirring at this temperature, water (35 kg) was slowly added and thesolution stirred for 2 h at 0-10° C. Then, 3% aqueous NaHCO3 (900 kg)and 1% aqueous NaOH (582 kg) was added. Next, NaBO₃.4H₂O (115.6 kg, 751mol) was added portion-wise over 1 h at 0-10° C. After stirring thereaction overnight at room temperature, additional NaBO₃.4H₂O (25.7 kg,167 mol) was added portion-wise over 1 h at 0-10° C. The reaction wasthen stirred an additional 6 h at room temperature.

The reaction was then extracted with ethyl acetate (230 kg) and theresulting organics washed with 3% aqueous NaHCO₃ (500 kg), followed bybrine (376 kg). The combined aqueous layers were further extracted withethyl acetate (2×325 kg). The organics were then treated with activatedcarbon (14.4 kg) for 2 h at 50-60° C. After filtration, the organicswere then concentrated and solvent switched to n-heptane to form acrystalline slurry. This slurry was then filtered and the cake waswashed with n-heptane. This wet cake was then dissolved in ethyl acetate(99 kg) at 50-60° C. n-Heptane (251 kg) was then added and the batchcooled to 0° C. The resulting slurry was then filtered and the cakewashed with n-heptane. The solids were then dried at 40-50° C. undervacuum to give tert-butyl[(2R,3S)-2-(2,5-difluorophenyl)-5-hydroxytetrahydro-2H-pyran-3-yl]carbamate.

Step L: tert-Butyl[(2R,3R)-2-(2,5-difluorophenyl)-5-hydroxytetrahydro-2H-pyran-3-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step K.

Step M: tert-Butyl[(2S,3R)-2-(2,5-difluorophenyl)-5-hydroxytetrahydro-2H-pyran-3-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step L.

Step N: tert-Butyl[(2S,3S)-2-(2,5-difluorophenyl)-5-hydroxytetrahydro-2H-pyran-3-yl]carbamate

This compound was made by following the same method described inIntermediate 1, Step M.

Step O: tert-Butyl[(2R,3S)-2-(2,5-difluorophenyl)-5-oxotetrahydro-2H-pyran-3-yl]carbamate

To 46.8 kg (142 mol) of tert-butyl[(2R,3S)-2-(2,5-difluorophenyl)-5-hydroxytetrahydro-2H-pyran-3-yl]carbamatein a stirred vessel was added acetonitrile (150 kg), acetic acid (50kg), and water (25 kg). After dissolving at room temperature, thesolution was cooled to 0° C. and RuCl₃.3H₂O (250 g, 956 mmol) in water(50 kg) was added under nitrogen. Then, NaBrO₃ (11.7 kg, 77.5 mol) wasadded in six portions every 1.5 h under nitrogen. After stirring at 0°C. for 6 h, 2-propanol (31 kg) was added over 30 min. at 0° C. Then,water (720 kg) was added at this temperature over 5 h. The resultingslurry was stirred overnight, filtered, and cake washed with water. Thesolids were then dried under vacuum at 40-60° C. to give tert-butyl[(2R,3S)-2-(2,5-difluorophenyl)-5-oxotetrahydro-2H-pyran-3-yl]carbamate.

INTERMEDIATE 2

2-(methylsulfonyl)-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-5-iumbenzenesulfonate Step A: tert-Butyl(3Z)-3-[(dimethylamino)methylene]-4-oxopyrrolidine-1-carboxylate

A solution of tert-butyl 3-oxopyrrolidine-1-carboxylate (53.4 kg, 288mol) in THF (133 kg) was treated with DMF-DMA (103 kg, 864 mol) in THF(472 kg) and heated at 65-70° C. under nitrogen for 20 h. The solutionwas cooled, evaporated under reduced pressure and solvent switched underdistillation to cyclohexane. The resulting slurry was then filtered,cake washed with cyclohexane, and then water. The solids were then driedunder vacuum at 35-40° C. to give tert-butyl(3Z)-3-[(dimethylamino)methylene]-4-oxopyrrolidine-1-carboxylate.

Step B: tert-Butyl6a-hydroxy-3a,4,6,6a-tetrahydropyrro[3,4-c]pyrazole-5(1H)-carboxylate

To a solution of tert-butyl(3Z)-3-[(dimethylamino)methylene]-4-oxopyrrolidine-1-carboxylate (58.2kg, 242 mol) in toluene (251 kg) at 35-45° C. was added hydrazinehydrate (14.6 kg, 290 mol) via drop-wise addition over 2 h. The mixturewas then stirred for 10 h at this temperature. The batch was then cooledto 0-10° C. and the slurry stirred for 6 h. This slurry was thenfiltered and the cake washed with n-heptane. The solids were then driedunder vacuum overnight at 35-50° C. to give tert-butyl6a-hydroxy-3a,4,6,6a-tetrahydropyrrol[3,4-c]pyrazole-5(1H)-carboxylate.

Step C: tert-Butyl 4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate

To a solution of tert-butyl6a-hydroxy-3a,4,6,6a-tetrahydropyrrol[3,4-c]pyrazole-5(1H)-carboxylate(47.0 kg, 207 mol) in dichloromethane (669 kg) at 0° C. was added amethanol solution of toluene-4-sulfonic acid monohydrate (3.7 kg, 20 molin 38 kg MeOH) drop-wise over 2 h. The reaction was then aged for 4 h atthis temperature. Then, 5% aqueous NaHCO₃ (91 kg) was added and stirredat room temperature for 30 min. The layers were then separated and theaqueous extracted with dichloromethane (312 kg). The combined organicswere washed with 5% brine (190 kg then 483 kg), treated with activatedcarbon (2.7 kg) and filtered. The resulting organics were dried withNa₂SO₄, filtered, and concentrated to 71-118 L. n-Heptane was then added(238 kg) and the batch further concentrated to 188-235 L. The slurry wascooled to 10-20° C., filtered, and the cake washed with n-heptane. Thesolids were dried under vacuum at 40-50° C. overnight to give tert-butyl4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate.

Step D: tert-Butyl2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate

A solution of tert-butyl4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate (30.0 kg, 143 mol)in 2-methyltetrahydrofuran (384 mg) was vacuum purged with nitrogenback-fill three times. The, triethylamine (25.0 kg, 247 mol) was addedand the batch cooled to −10-5° C. Then, methanesulfonyl chloride (21.4kg, 187 mol) was slowly added over 2 h. After stirring for 1 h at roomtemperature, water (150 kg) was added drop-wise at 5-15° C. This wasfollowed by addition of 1N HCl solution until the pH was 7. Theresulting layers were separated and the aqueous extracted with2-methyltetrahydrofuran (106 kg). The combined organics were washed withsaturated brine (2×150 kg), dried with Na₂SO₄, filtered, andconcentrated to 60-90 L.

The resulting crude was dissolved in 2-methyltetrahydrofuran (381 kg)and charged with a solution of potassium tert-butoxide in THF (805 g in6.6 kg THF). After stirring 1 h at room temperature under nitrogen, morepotassium tert-butoxide in THF (329 g in 3.0 kg THF) was added andstirred for 1 h. Analytical analysis indicates that tert-butyl2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylateis the major regioisomer, so saturated brine (154 kg) was then added.After brief agitation, the layers are separated and the oragnics arewashed with saturated brine (2×155 kg). The combined aqueous wastelayers were then extracted with 2-methyltetrahydrofuran (103 kg). Thecombined organics were treated with activated carbon (8.75 kg),filtered, and dried with Na₂SO₄. This was then filtered and concentratedto 60-90 L. This slurry was then heated to dissolve solids at 40-50° C.and n-heptane was added (34 kg). After cooling to room temperature for2-4 h, n-heptane (156 kg) was added and the slurry was then aged for 2-4h at 0-5° C. The slurry was filtered and the cake washed with n-heptane.The solids were dried under vacuum at 45-55° C. to give tert-butyl2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate.

Step E: tert-Butyl1-(methylsulfonyl)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxylate

This compound was made by following the same method described inIntermediate 1, Step D.

Step F: 2-(methylsulfonyl)-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-5-iumbenzenesulfonate

To a solution of tert-butyl2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate(32.1 kg, 111 mol) in iso-propylacetate (289 kg) was addedbenzenesulfonic acid (35.35 kg, 223 mol). The reaction was stirred for 3days at room temperature and then cooled to 0-10° C. and stirred anadditional 1 h. The resulting slurry was filtered and the cake washedwith iso-propylacetate. The solids were dried overnight under vacuum atroom temperature to give2-(methylsulfonyl)-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-5-iumbenzenesulfonate.

(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine

Step A: tert-Butyl{(2R,3S,5R)-2-(2,5-difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolol[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-yl}carbamate

A vessel was charged with N,N-dimethylacetamide (520.6 kg),2-(methylsulfonyl)-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazol-5-iumbenzenesulfonate (intermediate 2, 30.0 kg, 86.8 mol), and tert-butyl[(2R,3S)-2-(2,5-difluorophenyl)-5-oxotetrahydro-2H-pyran-3-yl]carbamate(intermediate 1, 31.2 kg, 95.3 mol). After dissolving at roomtemperature, the solution was cooled to 0-10° C. and sodiumtriacetoxyborohydride (24 kg, 113 mol) was added in four equal portionsevery 40 min. The reaction was then allowed to warm to room temperatureand stirred an additional 5 h. The solution was then cooled to 5-15° C.and water (672 kg) was added over 1-2 h. The resulting slurry wasfiltered and the cake washed sequentially with N,N-dimethylacetamide,twice with water, and then n-heptane. The solids were dried under vacuumat 40-60° C. to give tert-butyl{(2R,3S,5R)-2-(2,5-difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-yl}carbamate.

Step B:(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine

Benzenesulfonic acid (32.95 kg, 271 mol) was dissolved indichloromethane (1020 kg) under nitrogen. Then, 880 g of water was addedsuch that the solution KF was 0.2%. Next, tert-butyl{(2R,3S,5R)-2-(2,5-difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-yl}carbamate(38.4 kg, 100 mol) was added in three equal portions over 30 min. Thereaction was then aged overnight at room temperature. Next, water (733kg) was added over 1 h and the reaction stirred rapidly for 1 h. Thelayers were then separated, discarding the resulting organics layer. Tothe aqueous layer was charged dichloromethane (510 kg) followed bytriethylamine (22.4 kg, 592 mol). After agitation, the layers wereseparated and the aqueous extracted with dichloromethane (510 kg). Thecombined organics were washed with 7% aqueous NaHCO₃ (2×410 kg) and 5%brine (386 kg). The organics were then dried with Na₂SO₄, filtered, andtreated with activated carbon (6.2 kg of C-941). The carbon was filteredoff and the filtrate was concentrated under vacuum to 154-193 L. Thissolution was then warmed to 30-35° C. to dissolve solids (additionaldichloromethane may be added to dissolve solids). Next,iso-propylacetate (338 kg) was added and the solution stirred at roomtemperature for 1.5 h. Then, n-heptane (159 kg) was charged to thevessel drop-wise and stirred for 3 h. The slurry was then filtered andthe cake washed with n-heptane. This wet cake was then recrystallizedagain by dissolving it into dichloromethane and adding iso-propylacetateand n-heptane as before, filtering, and washing with n-heptane. Thesolids were dried under vacuum at 40-50° C. overnight to givecrystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-aminewas washed with cold 2:1EtOAc/hexanes to give the title compound as anoff-white solid. ¹H NMR (500 MHz, CD3OD): 1.71 (q, 1H, J=12 Hz),2.56-2.61 (m, 1H), 3.11-3.18 (m, 1H), 3.36-3.40 (m, 1H), 3.48 (t, 1H,J=12 Hz), 3.88-3.94 (m, 4H), 4.30-4.35 (m, 1H), 4.53 (d, 1H, J=12 Hz),7.14-7.23 (m, 2H), 7.26-7.30 (m, 1H), 7.88 (s, 1H). LC-MS: 399.04 [M+1].

Form I

Form I was produced by direct crystallization of the amorphous free baseof Compound I in ethyl acetate. The characterization results for XRPD,ssNMR, DSC, TGA and IR are shown below.

Form II

Crystalline Form II was produced by re-crystallization of Form I inisopropyl acetate and heptane 1:1 at room temperature. Form II wascharacterized using XRPD, ssNMR, DSC, TGA and IR. Conversion of Form IIinto Form I is slow but observed in all turnover experiments with 50-50seed including DCM-Heptane 25° C. over two days, IPAc 25° C. 17 hr, IPAc60° C. for one day, H₂O 60° C. over two weeks, three days, NMP-water 1-135° C. over three days. The relationship between Form I and Form II isenantiotropic having Form I as the most stable phase above 13° C.

Form III

Form III was produced by dissolving Form I in MeOH and evaporating thesolvent, followed by heating to 140° C. and isothermal for 10 min. Thisphase is metastable to Form I and II and its characterization waslimited to the amount of sample available. Form III was analyzed by XRPDand DSC.

Form IV

Form IV was produced by dissolving Form I in 1:1 THF-water andevaporating the solvent. Anhydrous Form IV is metastable to Form I andII and therefore the characterization was limited to the amount ofsample available. Form IV was analyzed using XRPD, DSC and TGA.

X-Ray Powder Diffraction

X-ray powder diffraction studies are widely used to characterizemolecular structures, crystalinity, and polymorphism. The X-ray powderdiffraction patterns for the solid phases for crystalline forms ofCompound I were generated on a Philips Analytical X'Pert PRO X-rayDiffraction System with PW3040/60 console. A PW3373/00 ceramic Cu LEFX-ray tube K-Alpha radiation was used as the source. The diffractionpeak positions were referenced by silicon (internal standard) which hasa 2 theta value of 28.443 degree. The experiments were analyzed atambient condition.

The crystalline forms described herein have a phase purity of at leastabout 5% of the form with the above X-ray powder diffraction and DSCphysical characteristics. In one embodiment the phase purity is at leastabout 10% of the form with the above solid-state physicalcharacteristics. In a second embodiment the phase purity is at leastabout 25% of the form with the above solid-state physicalcharacteristics. In a third embodiment the phase purity is at leastabout 50% of the form with the above solid-state physicalcharacteristics. In a fourth embodiment the phase purity is at leastabout 75% of the form with the above solid-state physicalcharacteristics. In a fifth embodiment the phase purity is at leastabout 90% of the form with the above solid-state physicalcharacteristics. In a sixth embodiment the crystalline forms of thepresent invention are the substantially phase pure forms with the abovesolid-state physical characteristics. By the term “phase purity” ismeant the solid state purity of the particular form with regard to aparticular crystalline form as determined by the solid-state physicalmethods described in the present application.

FIG. 1 is the X-ray powder diffraction (XRPD) pattern for Form I ofCompound I with selected d-spacings listed in Table 1.

TABLE 1 XRPD: Form I of Compound I 2θ(2 theta)(degrees) d-spacing (Å)10.3 8.63 12.7 6.99 14.6 6.07 16.1 5.51 17.8 4.97 19.2 4.61 22.2 4.0124.1 3.70 26.9 3.31

Crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Form I) is characterized by having at least four peaks in its powderX-ray diffraction pattern selected from the group consisting of 10.3±0.12θ, 12.7±0.1 2θ, 14.6±0.1 2θ, 16.1±0.1 2θ, 17.8±0.1 2θ, 19.2±0.1 2θ,22.2±0.1 2θ, 24.1±0.1 2θ and 26.9±0.1 2θ. The crystalline Form 1 can becharacterized by the following four peaks in its powder X-raydiffraction pattern 17.8±0.1 2θ, 19.2±0.1 2θ, 22.2±0.1 2θ and 24.1±0.12θ. The crystalline Form 1 can be characterized by the following fourpeaks in its powder X-ray diffraction pattern of FIG. 3.

FIG. 6 is the X-ray powder diffraction (XRPD) pattern for Form II ofCompound I with selected d-spacings listed in Table 2.

TABLE 2 X-ray powder diffraction: Form II of Compound I 2θ(2theta)(degrees) d-spacing (Å) 7.5 11.81 15.0 5.91 16.2 5.49 20.9 4.2522.0 4.04 27.0 3.30 27.6 3.24 33.3 2.69

Crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Form II) can be characterized by having at least four peaks in itspowder X-ray diffraction pattern selected from the group consisting of7.5±0.1 2θ, 15.0±0.1 2θ, 16.2±0.1 2θ, 20.9±0.1 2θ, 22.0±0.1 2θ, 27.0±0.12θ, 27.6±0.1 2θ, 33.3±0.1 2θ. The crystalline Form II can becharacterized by the following four peaks in its powder X-raydiffraction pattern 20.9±0.1 2θ, 22.0±0.1 2θ, 27.0±0.1 2θ and 27.6±0.12θ. Crystalline Form II of can be characterized by the X-ray powderdiffraction pattern of FIG. 6.

FIG. 11 is the X-ray powder diffraction (XRPD) pattern for Form III ofCompound I with selected d-spacings listed in Table 3.

TABLE 3 X-ray powder diffraction: Form III of Compound I 2θ(2theta)(degrees) d-spacing (Å) 14.5 6.09 15.9 5.58 17.3 5.11 18.7 4.7619.5 4.56 21.2 4.19 22.0 4.05 23.2 3.83

Crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Form III) can be characterized by having at least four peaks in itspowder X-ray diffraction pattern selected from the group consisting of14.5 +0.1 2θ, 15.9 +0.1 2θ, 17.3 +0.1 2θ, 18.7 +0.1 2θ, 19.5 +0.1 2θ,21.2 +0.1 2θ, 22.0 +0.1 2θand 23.2 +0.1 2θ. Crystalline Form III can becharacterized by the following four peaks in its powder X-raydiffraction pattern 19.5 +0.1 2θ, 21.2 +0.1 2θ, 22.0 +0.1 2θand 23.2+0.1 2θ. Crystalline Form III can be characterized by the X-ray powderdiffraction pattern of FIG. 11.

FIG. 14 is the X-ray powder diffraction (XRPD) pattern for Form IV ofCompound I with selected d-spacings listed in Table 4.

TABLE 4 X-ray powder diffraction: anhydrous Form IV of Compound I 2θ(2theta)(degrees) d-spacing (Å) 8.1 10.98 10.6 8.33 16.0 5.55 16.9 5.2419.5 4.56 21.3 4.18 23.3 3.82

Crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amine(Form IV) can be characterized by having at least four peaks in itspowder X-ray diffraction pattern selected from the group consisting of8.1±0.1 2θ, 10.6±0.1 2θ, 16.0±0.1 2θ, 16.9±0.1 2θ, 19.5±0.1 2θ, 21.3±0.12θ, 23.3±0.1 2θ and 25.4±0.1 2θ. Crystalline Form IV can becharacterized by the following four peaks in its powder X-raydiffraction pattern 16.9±0.1 2θ, 19.5±0.1 2θ, 21.3±0.1 2θ and 23.3±0.12θ. Crystalline Form IV can be characterized by the X-ray powderdiffraction pattern of FIG. 14.

ssNMR Spectra

Solid-state carbon-13 nuclear magnetic resonance spectrum was recordedon a Bruker AV400 NMR spectrometer using a Bruker 4 mm H/F/X BB doubleresonance CPMAS probe. The spectrum were collected utilizingproton/carbon-13 variable-amplitude cross-polarization (VACP) at 10 kHz,with a contact time of 3 ms. Other experimental parameters used for dataacquisition were a proton 90-degree pulse of 100 kHz, SPINAL64decoupling at 100 kHz, a pulse delay of 5 s, and signal averaging for1024 scans. The magic-angle spinning (MAS) rate was set to 10 kHz. ALorentzian line broadening of 10 Hz was applied to the spectrum beforeFourier Transformation. Chemical shifts are reported on the TMS scaleusing the carbonyl carbon of glycine (176.70 ppm.) as a secondaryreference.

Crystalline Form I can further characterized by the nuclear magneticresonance (NMR) spectra of FIG. 4. FIG. 4 is the ssNMR spectra for FormI of Compound I with selected peaks listed in Table 5.

TABLE 5 Selected ssNMR peaks for Form I of Compound I Peak (ppm)Relative Intensity 124.3 100 42.6 91 119.0 67 48.6 56 128.9 53 90.1 5073.2 46 163.6 44 59.9 42 157.9 38

Crystalline Form II can be further characterized by the nuclear magneticresonance (NMR) spectra of FIG. 9. FIG. 9 is the ssNMR spectra for FormII of Compound I with selected peaks listed in Table 6.

TABLE 6 Selected ssNMR peaks for Form II of Compound I Peak (ppm)Relative Intensity 116.9 100 127.5 82 42.2 78 132.1 61 73.5 60 79.0 5962.3 57 165.3 57 53.0 56 56.3 56IR Spectra

The Infrared spectrum was obtained using Attenuated Total Reflectance(ATR). The sample was placed directly onto the ATR-FTIR sampling deviceand the infrared spectrum was recorded using a Nicolet Nexus 670 FTIRspectrometer.

FIG. 5 is an IR spectra of Form I of Compound I. Crystalline Form I canbe further characterized by the IR spectra of FIG. 5.

FIG. 10 is an IR spectra of Form II of Compound I. Crystalline Form IIcan be further characterized by the IR spectra of FIG. 10.

In addition to the X-ray powder diffraction patterns described above,the crystalline forms of Compound I of the present invention werefurther characterized by means of their differential scanningcalorimetry (DSC) curves and their thermogravimetric analysis (TGA)curves.

DSC

Differential Scanning calorimetry data were acquired using TAInstruments DSC 2910 or DSC2000. Between 2 and 6 mg sample was weighedinto a pan and covered. This pan was then covered and placed at thesample position in the calorimeter cell. An empty pan is placed at thereference position. The calorimeter cell is closed and a flow ofnitrogen is passed through the cell. The heating program is set to heatthe sample at a heating rate of 10° C./min to a temperature ofapproximately 250° C. The data was analyzed using Universal Analysis2000 Version 3.9A. The thermal events were integrated between baselinetemperature points that are above and below the temperature range overwhich the thermal event is observed. The data reported are the onsettemperature, peak temperature and enthalpy.

Crystalline Form I can be further characterized by the differentialscanning calorimetric (DSC) curve of FIG. 3. Crystalline Form II can befurther characterized by the differential scanning calorimetric (DSC)curve of FIG. 8. Crystalline Form III can be further characterized bythe differential scanning calorimetric (DSC) curve of FIG. 13.Crystalline Form IV can be further characterized by the differentialscanning calorimetric (DSC) curve of FIG. 16.

TGA

Thermogravimetric data was acquired using a Perkin Elmer model TGA 7.Experiments were performed under a flow of nitrogen and using a heatingrate of 10° C./min to a maximum temperature of approximately 250° C.After automatically taring the balance, 5 to 20 mg of sample was addedto the platinum pan, the furnace was raised, and the heating programstarted. Weight/temperature data are collected automatically by theinstrument. Analyses of the results were carried out by selecting theDelta Y function within the instrument software and choosing thetemperatures between which the weight loss is to be calculated. Weightlosses are reported up to the onset of decomposition/evaporation.Crystalline Form I can be further characterized by the thermogravimetricanalysis (TGA) curve of FIG. 2. Crystalline Form II can be furthercharacterized by the thermogravimetric analysis (TGA) curve of FIG. 7.Crystalline Form III can be further characterized by thethermogravimetric analysis (TGA) curve of FIG. 12. Crystalline Form IVcan be further characterized by the thermogravimetric analysis (TGA)curve of FIG. 15.

A representative sample of Form I was analyzed by DSC and TGA accordingto the methods described above. Form I displays one endotherm (meltingof Form I confirmed by hot stage microscopy) with Tonset=173.48° C.,Tpeak=175.32° C., and ΔH=82.28 J/g (FIG. 3). Thermogravimetric analysisexhibits insignificant weight loss between room temperature and meltingpoint of Form I (FIG. 2).

A representative sample of Form II was analyzed by DSC (FIG. 8) and TGA(FIG. 7) according to the methods described above. The first endothermin the DSC curve is associated with the melting of Form II withT_(onset)=144.75° C., T_(peak)=147.59° C., and ΔH=23.41 J/g (FIG. 11).The first endotherm is followed by a recrystallization event to produceForm I at ˜150° C. and finally by the melting of form I atT_(onset)=170.18° C., T_(peak)=172.95° C., and ΔH=57.45 J/g. TG analysisexhibits minimum weight loss (trapped solvent) between room temperatureand melting of Form I.

DSC of Form III (FIG. 13) displays one endotherm associated with themelting of Form III with Tonset=164.30° C., Tpeak=169.38° C., andΔH=23.41 J/g. Thermogravimetric analysis (FIG. 12) shows ˜1% w/wresidual solvent in the initial material which was removed by heating at140 C and holding for 10 min.

DSC of Form IV (FIG. 16) displays one endotherm associated with themelting of Form IV with Tonset=171.25° C., Tpeak=172.30° C., andΔH=84.64 J/g. Less than 1% weight loss is observed up to melting usingTGA (FIG. 15).

What is claimed is:
 1. Crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(41i)-yl]tetrahydro-2H-pyran-3-amineof Compound I:

characterized by a solid-state carbon-13 nuclear magnetic resonance(NMR) spectra having chemical shifts at 163.6, 157.9, 128.9, 124.3 and119.0 ppm.
 2. The crystalline form of claim 1, characterized by asolid-state carbon-13 nuclear magnetic resonance (NMR) spectra havingchemical shifts at 163.6, 157.9, 128.9, 124.3, 119.0, 90.1, 73.2, 59.9,48.6 and 42.6 ppm.
 3. The crystalline form of claim 1, furthercharacterized by differential scanning calorimetric (DSC) thermogramwhich shows a maximum endothermal peak at or about 175.32° C.
 4. Thecrystalline form of claim 1, further characterized by athermogravimetric analysis (TGA) thermogram characterized byinsignificant weightloss between room temperature and melting point. 5.Crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(41i)-yl]tetrahydro-2H-pyran-3-amineof Compound I:

characterized by a solid-state carbon-13 nuclear magnetic resonance(NMR) spectra having chemical shifts at 132.1, 127.5, 116.9, 73.5 and42.2 ppm.
 6. The crystalline form of claim 5, characterized by asolid-state carbon-13 nuclear magnetic resonance (NMR) spectra havingchemical shifts at 132.1, 127.5, 116.9, 79.0, 73.5, 62.3, 56.3, 53.0 and42.2 ppm.
 7. The crystalline form of claim 5, further characterized bythe differential scanning calorimetric (DSC) thermogram which shows anendothermal peak at or about 172.95° C.
 8. Crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amineof Compound I:

characterized by having at least four peaks in its powder X-raydiffraction pattern selected from the group consisting of 14.5 +0.1 2θ,15.9 +0.1 2θ, 17.3 +0.1 2θ, 18.7 +0.1 2θ, 19.5 +0.1 2θ, 21.2 +0.1 2θ,22.0 +0.1 2θ and 23.2 +0.1 2θ and further characterized by differentialscanning calorimetric (DSC) thermogram which shows an endothermal peakat or about 169.38° C.
 9. Crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amineof Compound I:

characterized by having at least four peaks in its powder X-raydiffraction pattern selected from the group consisting of 8.1 +0.1 2θ,10.6 +0.1 2θ, 16.0 +0.1 2θ, 16.9 +0.1 2θ, 19.5 +0.1 2θ, 21.3 +0.1 2θ,23.3 +0.1 2θ and 25.4 +0.1 2θ and further characterized by differentialscanning calorimetric (DSC) thermogram which shows an endothermal peakat or about 172.30° C.
 10. A method of treating Type 2 diabetescomprising administering to a mammal in need of such treatment atherapeutically effective amount of a crystalline form according toclaim
 1. 11. A pharmaceutical composition comprising a drug substancethat comprises crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amineof claim 1 and a pharmaceutically acceptable carrier.
 12. A method oftreating Type 2 diabetes comprising administering to a mammal in need ofsuch treatment a therapeutically effective amount of a crystalline formaccording to claim
 2. 13. A pharmaceutical composition comprising a drugsubstance that comprises crystalline(2R,3S,5R)-2-(2,5-Difluorophenyl)-5-[2-(methylsulfonyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl]tetrahydro-2H-pyran-3-amineof claim 2 and a pharmaceutically acceptable carrier.